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This paper describes a simple and inexpensive procedure based on the use of clove oil as a tissue\u2010clearing, staining, and mounting medium that helps overcome the problem of slow dye penetration and tissue opaqueness and allows muscles and several other organ systems to be visualized by confocal or epifluorescent microscopy. This clove oil\u2010induced fluorescence (COIF) method relies on the ability of clove oil to accumulate in muscles and some other tissues and become steadily fluorescent if irradiated at 488\u2009nm. For this method, animals were fixed in 70% ethanol or 4% formaldehyde, then dehydrated and mounted in clove oil. Heavily pigmented animals were additionally bleached in hydrogen peroxide prior to the dehydration step. The COIF method showed excellent results in all major groups of arthropods and some mollusks and annelids revealing the three\u2010dimensional arrangement of muscles, gonads, glands, cellular nuclei and some parts of the nervous and digestive systems. In the other animal groups tested, clove oil stained all tissues making it difficult to observe the anatomical details. The COIF method is advantageous in some respects over other methods such as phalloidin staining because of its tissue penetration and clearing abilities, low cost of the reagents, resistance to photodamage and the possibility of staining museum specimens.<\/jats:p>","DOI":"10.1002\/jemt.24295","type":"journal-article","created":{"date-parts":[[2023,1,27]],"date-time":"2023-01-27T09:34:30Z","timestamp":1674812070000},"page":"565-572","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["A simple and low\u2010cost method to visualize musculature and other aspects of anatomy by confocal microscopy"],"prefix":"10.1002","volume":"86","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3010-8341","authenticated-orcid":false,"given":"Anatoly A.","family":"Petrov","sequence":"first","affiliation":[{"name":"Zoological Institute, Russian Academy of Sciences 1 Universitetskaya Embankment  St. Petersburg Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0602-0913","authenticated-orcid":false,"given":"Elena V.","family":"Soldatenko","sequence":"additional","affiliation":[{"name":"Zoological Institute, Russian Academy of Sciences 1 Universitetskaya Embankment  St. Petersburg Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2023,1,27]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.als.2016.05.001"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.proghi.2016.04.001"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1002\/jmor.10789"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.3791\/50664"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1101\/pdb.prot4525"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.3390\/molecules26216387"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2016.162"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jneumeth.2008.04.031"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1080\/00779962.1978.9722313"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jcz.2017.09.001"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.7717\/peerj.7251"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2015.06.067"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.1117\/1.JBO.21.8.081205"},{"key":"e_1_2_9_15_1","first-page":"139","article-title":"A modified technique for the preparation of specimens of Sternorryncha for taxonomic studies","volume":"24","author":"Sirisena U.","year":"2013","journal-title":"Tropical Agricultural Research"},{"key":"e_1_2_9_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.freeradbiomed.2017.03.014"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.asd.2013.12.006"},{"key":"e_1_2_9_18_1","doi-asserted-by":"publisher","DOI":"10.1111\/joa.13309"},{"key":"e_1_2_9_19_1","doi-asserted-by":"publisher","DOI":"10.5281\/zenodo.4641412"}],"container-title":["Microscopy Research and Technique"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jemt.24295","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/jemt.24295","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jemt.24295","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,16]],"date-time":"2025-10-16T13:43:05Z","timestamp":1760622185000},"score":1,"resource":{"primary":{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/10.1002\/jemt.24295"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,1,27]]},"references-count":18,"journal-issue":{"issue":"5","published-print":{"date-parts":[[2023,5]]}},"alternative-id":["10.1002\/jemt.24295"],"URL":"https:\/\/doi.org\/10.1002\/jemt.24295","archive":["Portico"],"relation":{},"ISSN":["1059-910X","1097-0029"],"issn-type":[{"value":"1059-910X","type":"print"},{"value":"1097-0029","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,1,27]]},"assertion":[{"value":"2022-10-26","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2023-01-13","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2023-01-27","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}